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目的建立了使用聚四氟乙烯带盖水解管,以NaOH溶液为水解液的碱解法测定动物水解蛋白中L-羟脯氨酸(L-Hydroxyproline,L-Hyp)含量。方法将样品置于10 ml水解管中,加入6 ml 2.5 mol/L NaOH溶液在110℃烘箱中加热2 h,水解出的L-Hyp经氯胺T氧化后与对二甲氨基苯甲醛反应生成红色络合物,在(558±2)nm处测定其吸光度。结果在优化实验条件下,该方法的线性范围0～10μg/ml(r=0.999 3),检出限1.35μg/g,样品测定的RSD在1.0%～2.3%,加标回收率为88.7%～96%(加标量为30 mg/kg)。结论相对酸解法,碱解法的水解效率和酸解法基本一致,样品前处理操作简单,缩短了测定时间,灵敏度、重复性和稳定性良好。
OBJECTIVE To establish a method for determination of L-Hydroxyproline (L-Hydroxyproline, L-Hydroxyproline, L-Hydroxyproline, L-Hydroxyproline, Methods The sample was placed in a 10 ml hydrolysis tube and heated in an oven at 110 ° C for 6 h by adding 6 ml of 2.5 mol / L NaOH solution. The hydrolyzed L-Hyp was oxidized by chloramine T and reacted with p-dimethylaminobenzaldehyde Red complex, absorbance was measured at (558 ± 2) nm. Results Under the optimized experimental conditions, the linear range of this method was 0 ~ 10μg / ml (r = 0.999 3), the detection limit was 1.35μg / g, the RSD of the sample was 1.0% ~ 2.3% and the recovery was 88.7% ~ 96% (plus standard 30 mg / kg). Conclusion Compared with acid hydrolysis method, the hydrolysis efficiency and acid hydrolysis method of alkali hydrolysis are basically the same. The pretreatment of sample is simple and the measurement time is shortened. The sensitivity, repeatability and stability are good.